US5045446A - Lyophilization of cells - Google Patents

Lyophilization of cells Download PDF

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US5045446A
US5045446A US07378349 US37834989A US5045446A US 5045446 A US5045446 A US 5045446A US 07378349 US07378349 US 07378349 US 37834989 A US37834989 A US 37834989A US 5045446 A US5045446 A US 5045446A
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cells
process
solution
polymer
monosaccharide
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Raymond P. Goodrich, Jr.
Christine M. Williams
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Cobe Laboratories Inc
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Cryopharm Corp
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    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01NPRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES, AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N1/00Preservation of bodies of humans or animals, or parts thereof
    • A01N1/02Preservation of living parts
    • A01N1/0205Chemical aspects
    • A01N1/021Preservation or perfusion media, liquids, solids or gases used in the preservation of cells, tissue, organs or bodily fluids
    • A01N1/0221Freeze-process protecting agents, i.e. substances protecting cells from effects of the physical process, e.g. cryoprotectants, osmolarity regulators like oncotic agents
    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01NPRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES, AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N1/00Preservation of bodies of humans or animals, or parts thereof
    • A01N1/02Preservation of living parts
    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01NPRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES, AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N1/00Preservation of bodies of humans or animals, or parts thereof
    • A01N1/02Preservation of living parts
    • A01N1/0205Chemical aspects

Abstract

A process and medium are disclosed for the lyophilization of cells which comprises the use of solutions including monosaccharide hexoses and pentoses, and/or biocompatible amphipathic polymers to permit the reconstitution of viable cells.

Description

FIELD OF THE INVENTION

This is a continuation-in part of Ser. No. 237,583, filed Aug. 25, 1988, now abandoned. This invention relates to the general field of biochemistry and medical sciences, and specifically to processes for the preservation, storage and reconstitution of cells, particularly red blood cells.

BACKGROUND AND SUMMARY OF THE INVENTION

Laboratory cell preservation and storage have been significant problems for a variety of plant and animal cells. Freezing the cells in an aqueous solution and thawing the cells prior to use is not uncommon, but the viability of the cells after this process is generally severely affected and chromosome abnormalities often result from this freeze-thaw process. In addition, the expense of keeping the cells frozen is significant.

For example, there has been a need for improved methods for the storage of blood and blood constituents. Blood is a major tissue of the human body, and has as a predominant role the delivery of oxygen from the lungs to peripheral tissues. This role is carried out by erythrocytes, i.e., red blood cells (RBC). The oxygen is furnished from the lungs by an exchange-diffusion system brought about by a red, iron-containing protein called hemoglobin. When hemoglobin combines with oxygen, oxyhemoglobin is formed and after oxygen is given up to the tissues, the oxyhemoglobin is reduced to deoxyhemoglobin.

The red cell membrane is composed of two major structural units, the membrane bilayer and a cytoskeleton. A lipid bilayer and integral membrane proteins form the membrane bilayer, which has little structural strength and fragments readily by vesiculation. The other major component, the membrane skeleton, stabilizes the membrane bilayer and provides resistance to deformation. The cytoskeleton is linked to the bilayer in the erythrocyte membrane, possibly by lipid-protein as well as protein-protein associations. The hemoglobin, and other RBC components, are contained within the red cell membrane.

In adults, bone marrow is active in the formation of new red blood cells. Once erythrocytes enter the blood, these cells have an average lifetime of about 120 days. In an average person, about 0.83% of the erythrocytes are destroyed each day by phagocytosis, hemolysis or mechanical damage in the body, and the depleted cells are renewed from the bone marrow.

A wide variety of injuries and medical procedures require the transfusion of whole blood or a variety of blood components. Every patient does not require whole blood and, in fact, the presence of all of the blood components can cause medical problems. Separate blood fractions can be stored under those special conditions best suited to assure their biological activity at the time of transfusion. For example, when donor blood is received at a processing center, erythrocytes are separated and stored by various methods. Such cells are storable in citrate-phosphate-dextrose at 4° C. for up to five weeks, generally as a unit of packed erythrocytes having a volume of from 200 to 300 ml and a hematocrit value (expressed as corpuscular volume percent) of 70 to 90. Erythrocytes may also be treated with glycerol and then frozen at from -30° to -196° C. and stored for up to seven years in a glycerol solution, but must be kept frozen at low temperatures in order to survive sufficiently for transfusion. Both these methods require careful maintenance of storage temperature to avoid disruption of the desired biological activity of the erythrocytes, and provide a twenty-four hour survival time for at least 70% of the transfused cells, which is considered to be an acceptable level for use in transfusion practice in accordance with the American Association of Blood Bank standards.

It has thus been a desideratum to obtain a method for the storage of cells, and in particular red blood cells, which is not dependent on the maintenance of specific storage temperatures or other storage conditions. Such a method would facilitate the availability of erythrocytes for medical purposes and assist in the storage and shipment of various mammalian cells and plant cells, particularly protoplasts, for research and hybrid development.

One such desired method has been the lyophilization (freeze-drying) of cells, since such cells could be stored at room temperature for an extended period of time and easily reconstituted for use. Freeze-dried red blood cells could thus be easily stored for use in transfusions. However, prior to our invention, it has been impossible to freeze-dry erythrocytes in a manner which permits the reconstitution of the cells to form erythrocytes with an intact cytoskeleton and biologically-active hemoglobin, i.e., viable red blood cells. When RBCs have been lyophilized according to previous methods, for example in either an aqueous or phosphate-buffered saline (PBS) solution, the reconstituted cells are damaged to the extent that the cells are not capable of metabolizing, and the cell hemoglobin cannot carry oxygen. Glutaraldehyde-fixed erythrocytes, which have been lyophilized and reconstituted, have found use primarily in agglutination assays.

The process of the present invention allows for the lyophilization of cells under conditions which are not deleterious to the structure and the biological activity of the cell, and which permits the reconstitution of the lyophilized cells to form cells which are identical to the natural cells in a biological or botanical activity. Briefly, the process comprises immersing a plurality of cells in an essentially isotonic aqueous solution containing a carbohydrate, and which preferably includes an amphipathic polymer, freezing the solution, and drying the solution to yield freeze-dried cells which, when reconstituted, produce a significant percentage of intact and viable cells.

While the invention is applicable to a wide variety of plant and animal cells, the process of the invention is preferably applied to red blood cells and allows for the lyophilization of erythrocytes under conditions which maintain structure of the cell and the biological activity of the hemoglobin, and which permits the reconstitution of the lyophilized red blood cells to allow use on a therapeutic level. The carbohydrate of the invention is biologically compatible with the cells, that is, non-disruptive to the cells, and is preferably one which permeates, or is capable of permeating, the membrane of the cells. The carbohydrate may be selected from the group consisting of monosaccharides, since disaccharides do not appear to permeate the membrane to any significant extent. Monosaccharide pentoses and hexoses are preferred in concentrations of from about 7.0 to 37.5%, preferably about 23%. Xylose, glucose, ribose, mannose and fructose are employed to particular advantage. The lyophilization of RBCs in such a carbohydrate solution improves the recovery after lyophilization to at least 50% intact cells, as opposed to the fusing and destruction of the cell membrane during lyophilization in water solutions without the carbohydrate. Such reconstituted cells are only useful in producing ghost cells for agglutination assays or biochemical research, i.e., as model membrane systems. They are not viable cells capable of transporting oxygen or metabolizing.

As stated above, the addition to the carbohydrate solution of a water soluble, biologically compatible polymer adds significantly to the percentage of biologically-active hemoglobin which is retained in the cells and recovered after reconstitution of red blood cells after lyophilization. The polymer will preferably be amphipathic, meaning that there are hydrophilic and hydrophobic portions on a single molecule of the polymer. The polymer may be present in the solution in concentrations of from 0.7% up to saturation. Preferably, the polymer has a molecular weight in the range of from about 1K to about 360K, most preferably from about 5K to 80K, and most preferably to 50K, and is present in a concentration of from about 3.5% up to the limit of solubility of the polymer in the solution. Polymers selected from the group consisting of polyvinylpyrrolidone (PVP) and polyvinylpyrrolidone derivatives, and dextran and dextran derivatives provide significant advantages. Amino acid based polymers (i.e., proteins) or hydroxyethyl starch may also be employed. Other amphipathic polymers may be used, such as poloxamers in any of their various forms. The use of the carbohydrate-polymer solution in the lyophilization of red blood cells allows for the recovery of intact cells, a significant percentage of which contain biologically-active hemoglobin. While not intending to be bound by any theory, the amphipathic properties of the polymer allow them to bind to the cell membrane while protecting the membrane surface by extension of the hydrophilic portion into the aqueous environment. This may alleviate the damage to the cell membrane which causes other problems, such as cell aggregation. The use of the carbohydrate-polymer solution in the lyophilization of red blood cells allows for the recovery of intact cells, a significant percentage of which contain biologically-active hemoglobin.

As is shown by the data set forth below, the described solutions provide media which permit cells, particularly red blood cells, to be subjected to the stresses of freezing, water sublimation and reconstitution and to form freeze-dried cells which may be reconstituted to yield cells which are capable of functioning normally.

Unless indicated otherwise by the terminology or the context, all percentages set forth herein are expressed as weight percentages (i.e., weight of the solute versus the total weight of the solution).

DESCRIPTION OF THE PREFERRED EMBODIMENT

As noted above, the process of the invention provides a medium for the lyophilization and reconstitution of intact and biologically-active erythrocytes. While the media of the invention are novel it will be understood that apparatus and related techniques are known by those of skill in the art for the lyophilization of various materials, and biological samples in particular, and only the specific temperatures and apparatus employed in the examples are described herein. From this description, one of ordinary skill in the art will be capable of employing the media of the invention in a process for the freeze-drying and reconstitution of intact, viable red blood cells.

The term lyophilization is broadly defined as freezing a substance and then reducing the concentration of one of the solutes, namely water, by sublimation and desorption, to levels which will no longer support biological or chemical reactions. Usually, the drying step is accomplished in a high vacuum. However, with respect to the storage of cells and particularly erythrocytes, the extent of drying (the amount of residual moisture) is of critical importance in the ability of cells to withstand long-term storage at room temperature. In the method of the invention, cells may be lyophilized to a residual water content of less than 10%, preferably less than 5%, and most preferably to a water content of less than 3%.

EXAMPLE ONE

Packed red blood cells of variable blood type were obtained from a hospital blood donor center or drawn from healthy volunteers using heparin as an anticoagulant.

Repeated samples of these blood cells were washed with a phosphate buffered saline solution (10 mM mono- and di-basic sodium phosphate, 150 mM sodium chloride, 5 mM dextrose, and 10 mM adenosine at pH 7.2) three times with centrifugation at 14,000 rpm for 6 to 10 seconds to separate plasma and/or other cell types from the red blood cells.

Samples of these packed red blood cells were then suspended in a lyophilizing buffer containing 26.5% glucose in PBS solution at pH 7.2.

The suspension was then transferred to a flask which was subsequently immersed in liquid nitrogen (-196° C.) until the sample was frozen. The flask was rotated evenly in the liquid nitrogen to assure even dispersion of solution on the walls of the flask.

The frozen samples were transferred to a bench top lyophilizer (Labconco model 4.5) operating at less than 100 microns of mercury vacuum with an inner chamber temperature of -56° C. Samples were allowed to dry thoroughly (6-24 hours) until crystalline in appearance and brittle to touch and the flask was allowed to return to room temperature.

The samples were rehydrated at 37° C. using a solution containing 25.5% sucrose in a phosphate buffered saline solution. A volume of the rehydrating solution was added equivalent to the initial volume of the sample prior to drying.

It was found upon examination of the cells with an optical microscope that about 50% of the red blood cells had intact cell membranes. However, the hemoglobin was found not to be cell associated. Nonetheless, the hemoglobin in the solution was functional and if present in the cells would be effective as an oxygen carrier. Repeating this procedure with fructose and ribose solutions having concentrations of from about 7.0 to 37.5% produced nearly equal results, as did buffered solutions of xylose and mannose at concentrations of from about 7.0 to 37.5%.

Specifically, various monosaccharides were employed in the lyophilization of RBCs as described in this example, and the distribution of the hemoglobin in the recovered solution was noted. Oxyhemoglobin is capable of transporting oxygen to mammalian tissue. Methemoglobin is hemoglobin which cannot bind oxygen, but can possibly be reversed to form oxyhemoglobin when found in lower concentrations y the enzyme NADH methemoglobin reductase. Hemochrome is irreversibly degraded hemoglobin. In Table I, the recovery of greater than 90% oxyhemoglobin from cells lyophilized with solutions of ribose, mannose, fructose, xylose and glucose is shown.

              TABLE I______________________________________% Carbohydrate       % OxyHb   % MetHb   % Hemochrome______________________________________23.1 Ribose 93.1      5.4       1.526.5 Mannose       94.2      6.0       026.5 Fructose       98.0      1.3       0.726.5 Sorbose       56.9      40.9      2.315.2 Galactose       81.0      17.3      1.723.1 Xylose 96.7      3.6       026.5 Glucose       98.1      1.8       0.1______________________________________
EXAMPLE TWO

A number od samples of packed red blood cells, obtained and washed as described in Example One, were suspended in a lyophilizing buffer containing 23.1% glucose and a concentration of either (a) 18% of 10K or (b) 12.8% of 40K polyvinylpyrrolidone in PBS at pH 7.2.

The suspension was then transferred to a flask which was subsequently immersed in liquid nitrogen (-196° C.) until the sample was frozen. The flask was rotated evenly in the liquid nitroqen to assure even dispersion of solution on the walls of the flask.

The frozen sample was transferred to a bench top lyophilizer (Labconco model 4.5) operating at less than 100 microns of mercury vacuum with an inner chamber temperature of -56° C. Samples were allowed to dry thoroughly (6-24 hours) until crystalline in appearance and brittle to touch and the flask was allowed to return to room temperature.

The samples were rehydrated at 37° C. using a solution containing 25.5% sucrose in a phosphate buffered saline solution. A volume of the rehydrating solution was added equivalent to the initial volume of the sample prior to drying.

The samples were centrifuged at 14,000 rpm in an Eppendorf microcentrifuge to pellet the rehydrated red blood cells in suspension.

The results of incorporating the polymer with the above described carbohydrate in the buffered lyophilizing solution produced surprising results not only in that the recovery of intact cells was maintained at 52.9±7.9%, but in addition the solution allowed hemoglobin retention by the cells of from 27.4 up to 42.2% for the 10K PVP and from 57.3 up to 65.5% for the 40K PVP, with greater than 80% of the hemoglobin being oxyhemoglobin. Further testing has shown that 24K PVP at a concentration of 12.8% and a glucose concentration of 23.1% produces even better results both in cell and hemoglobin recovery.

EXAMPLE THREE

The procedure described in Example Two was repeated, with different carbohydrates substituted for glucose in the lyophilizing buffer. Two different molecular weights of polyvinylpyrrolidone were used. The results are shown in Table II.

              TABLE II______________________________________PVPMW    % Carbohydrate              % Cell Recovery                           % Hb Recovery______________________________________10K*  12.2 Galactose              27.7         10.3 21.7 Mannose 57.6         30.6 18.8 Xylose  63.9         32.3 21.7 Fructose              54.6         28.1 21.7 Glucose 59.0         28.624K** 13.0 Galactose              26.3         13.8 23.1 Mannose 51.8         57.2 20.0 Xylose  48.4         55.9 23.1 Fructose              48.8         59.3 23.1 Glucose 59.0         52.7______________________________________ *with a PVP concentration of from 18.1 to 20.3%. **with a PVP concentration of from 12.8 to 14.5%.

Trehalose and sucrose in the lyophilizing solution showed marginal cell recovery, but no hemoglobin recovery.

EXAMPLE FOUR

The procedure described in Example Two (using from 21.7 to 26.3% glucose as the carbohydrate) was repeated substituting polyvinylpyrrolidone of different molecular weights and concentrations for those used in the lyophilizing buffer of the previously described Example. All other conditions were repeated as described in Example Two. The results are shown in Table III. The column headed MCHC refers to the mean cell hemoglobin content of the reconstituted cells. The MCHC of normal RBCs is 34±2. Table III demonstrates that PVP may be employed with molecular weights of from 10 to 40K in concentrations of from 0.7 to 18.1%. The 40KT PVP had a viscosity of about 26 to 35 poise, and the 40K PVP had a viscosity of about 28 to 32 poise. Maltose showed no cell or hemoglobin recovery.

              TABLE III______________________________________PVP    Conc.MW     (%)     % Hb Recoverv  MCHC______________________________________ 10K   3.5             13.6           --  6.8             15.0           34.9  12.8    30.1 ±                  4.1 (n = 3)                           20.9 ±                                 3.1 (n = 3)  18.1            36.5           28.1 24K   3.5             24.7           17.3  6.8             52.9           20.9  12.8    52.7 ±                  6.3 (n = 4)                           27.4 ±                                 4.3 (n = 4)  18.1    52.2 ±                  6.9 (n = 2)    -- 40K   3.5             17.7           --  6.8             31.0           22.5  12.8    61.4 ±                  4.1 (n = 3)                           25.7 ±                                 9.2 (n = 3)  18.1    52.0 ±                  1.7 (n = 2)    37.4 40KT  3.5             17.7           --  6.8             31.8           25.0  12.8    56.8 ±                  0.4 (n = 2)                           36.3 ±                                 2.8 (n = 2)  18.1            50.0           29.4360K   0.7              9.4           --  8.5             12.2           --______________________________________
EXAMPLE FIVE

The experiment described in Example Two was repeated using polymers other than polyvinylpyrrolidone in the lyophilizing buffer. The results are summarized in Table IV.

              TABLE IV______________________________________Polymer    MW        % Conc.  % Hb Recovery______________________________________Dextran    10K       3.5      26.1                6.8      29.8                12.8     26.5                18.1     30.2      40K       3.5      24.7                6.8      19.5                12.8     25.9                18.1     16.6      80K       3.5      15.2                6.8      26.5                12.8     20.2                18.1     18.7Ficoll     70K       3.5      17.3                6.8      19.1      400K      0.7      17.2                3.5      17.9Fish Gelatin         1.4      19.0                6.8      18.4Dextrin              1.4      20.4                6.8      13.1Albumin              1.4      29.7______________________________________
EXAMPLE SIX

Samples of packed red blood cells were obtained and washed as described in Example One. These cells were suspended in a lyophilizing buffer of 12.8% 24K PVP and 23.1% glucose in phosphate buffered saline. The samples were lyophilized and reconstituted as described in Example Two, but with the various solutions used in the reconstitution of the cells. When water was the sole reconstituting liquid, the cells lysed within thirty minutes after reconstitution. An isotonic reconstituting solution, such as PBAS or PBSGA (PBS with the addition of 5 mmol glucose and 10 mmol adenosine) showed improvement, as did the use of reverse PBS, which employs potassium rather than sodium salts. Significant improvements were shown by the use of concentrations of up to 12.8% of either 10K or 24K PVP in the reconstitution solution.

The use of a carbohydrate in a minimum concentration of at least 0.7 to 3.6%, and most preferably at least 3.6%, provides better cell morphology after reconstitution. Both mono- and disaccharides may be employed for this purpose, although glucose, mannose, trehalose and sucrose are preferred with sucrose being the most preferred carbohydrate. These data are shown in Table V, wherein all carbohydrate and polymer solutions are formed in PBS.

              TABLE V______________________________________       % Cell      % HbSolution    Recovery    Recovery  MCHC______________________________________Water       49.3 ± 3.0                   37.4 ± 1.1                             29.9 ± 1.8PBS         59.2        34.4      24.8PBSGA       60.6        42.4      31.2Reverse PBS 52.6        51.3      25.8Glucose 15.9%       52.5        57.3      32.9Mannose 15.9%       55.5        60.7      28.0Trehalose 27.4%       65.7        59.4      24.9Sucrose 1.7%       61.7        45.6      24.4 3.3%       43.8        46.2      27.3 7.9%       49.5        52.8      24.625.5%       49.6 ± 10.6                   51.4 ± 5.1                             25.5 ± 2.1 4.8% 10K PVP       55.6 ± 11                   52.3 ± 3.0                             23.5 ± 1.416.7% 10K PVP       60.8        67.7      28.4 4.8% 24K PVP       52.2        38.3      26.016.7% 24K PVP       53.8 ± 9.4                   73.1 ± 8.1                             28.2 ± 8.7 3.6% 10K PVP +       65.0 ± 6.5                   59.0 ± 7.6                             28.2 ± 8.724.6% Sucrose13.0% 10K PVP +       39.5        61.6      27.822.2% Sucrose 3.6% 24K PVP +       64.8        59.3 ± 6.9                             26.524.6% Sucrose13.0% 24K PVP +       77.7        76.4 ± 4.2                             31.522.2% Sucrose______________________________________

From the foregoing description, one skilled in the art can readily ascertain the essential characteristics of the invention and, without departing from the spirit and scope thereof, can adapt the invention to various usages and conditions. Changes in form and substitution of equivalents are contemplated as circumstances may suggest or render expedient, and although specific terms have been employed herein they are intended in a descriptive sense and not for purposes of limitation.

Claims (18)

What is claimed is:
1. A process for the lyophilization of cells having a cell membrane, comprising immersing a plurality of cells in a water solution which includes a monosaccharide, which is capable of permeating the membrane of the cells, in a concentration of from about 7.0 to 37.5%; freezing the solution; and drying the frozen cells by sublimation of the water.
2. The process of claim 1 wherein the monosaccharide is selected from the group consisting of pentoses and hexoses.
3. The process of claim 1 or 2 wherein the monosaccharide is selected from the group consisting of xylose, glucose, ribose, mannose and fructose.
4. The process of claim 3 wherein the monosaccharide is present in the solution in a concentration of about 23%.
5. A process for the lyophilization of cells, comprising:
immersing a plurality of cells in a buffered solution which includes:
a monosaccharide which is present in the solution in a concentration of from about 7.0 to 37.5%, and
a polymer having a molecular weight of from about 1K to about 360K which is present in a concentration of from about 0.7% up to saturation in the solution;
freezing the solution; and
drying the cells by sublimation of the water.
6. The process of claim 5 wherein said polymer is amphipathic.
7. The process of claim 5 wherein said polymer has a molecular weight in the range of about 5K to about 80K.
8. The process of claim 5, 6 or 7 wherein the monosaccharide is selected from the group consisting of pentoses and hexoses.
9. The process of claim 8 wherein the monosaccharide is selected from the group consisting of xylose, glucose, ribose, mannose and fructose.
10. The process of claim 8 wherein the polymer is selected from the group consisting of polyvinylpyrrolidone and dextran.
11. The process of claim 10 wherein the polymer is polyvinylpyrrolidone.
12. A process for the lyophilization of cells, comprising:
immersing a plurality of cells in a buffered solution which includes:
a monosaccharide which is present in the solution in a concentration of from about 7.0 to 37.5%, and
a polymer having a molecular weight of from about 1K to about 360K which is present in a concentration of from about 0.7% up to saturation in the solution;
freezing the solution; and
drying the cells by sublimation of the water.
13. The process of claim 12 wherein said polymer is amphipathic.
14. The process of claim 12 wherein said polymer has a molecular weight in the range of about 5K to about 80K.
15. The process of claim 12, 13 or 14 wherein the monosaccharide is selected from the group consisting of pentoses and hexoses.
16. The process of claim 15 wherein the monosaccharide is selected from the group consisting of xylose, glucose, ribose, mannose and fructose.
17. The process of claim 15 wherein the polymer is selected from the group consisting of polyvinylpyrrolidone and dextran.
18. The process of claim 17 wherein the polymer is polyvinylpyrrolidone.
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US07378349 US5045446A (en) 1988-08-26 1989-07-11 Lyophilization of cells
CA 609229 CA1327763C (en) 1988-08-25 1989-08-24 Lyophilization of cells
IL9141389A IL91413D0 (en) 1988-08-26 1989-08-24 Process and solution for lyophilization of cells
DE1989621945 DE68921945D1 (en) 1988-08-26 1989-08-25 A process for the lyophilization of cells Lyophilisierungsmedien and methods for recovery of lyophilized cells.
JP22011589A JP2809735B2 (en) 1988-08-26 1989-08-25 Freeze-drying method of cells
AT89308672T AT120335T (en) 1988-08-26 1989-08-25 A process for the lyophilization of cells lyophilisierungsmedien and method for restoring of lyophilized cells.
DE1989621945 DE68921945T2 (en) 1988-08-26 1989-08-25 A process for the lyophilization of cells Lyophilisierungsmedien and methods for recovery of lyophilized cells.
EP19890308672 EP0356257B1 (en) 1988-08-26 1989-08-25 Processes for the lyophilization of cells including lyophilization media, and methods of reconstituting lyophilized cells
EP19920110201 EP0508496A1 (en) 1988-08-26 1989-08-25 Methods of reconstituting lyophilized cells including reconstituting media
US07708147 US5425951A (en) 1988-05-18 1991-05-31 Method of reconstituting lyophilized cells
US08260165 US5958670A (en) 1988-05-18 1994-06-15 Method of freezing cells and cell-like materials
US08412305 US5648206A (en) 1988-08-26 1995-03-29 Lyophilization of cells
US08479975 US6007978A (en) 1988-05-18 1995-06-07 Method of freezing cells and cell-like materials

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1993014191A1 (en) * 1992-01-21 1993-07-22 Cryopharm Corporation Method of freezing cells and cell-like materials
US5336616A (en) * 1990-09-12 1994-08-09 Lifecell Corporation Method for processing and preserving collagen-based tissues for transplantation
US5340592A (en) * 1988-05-18 1994-08-23 Cobe Laboratories, Inc. Lyophilization of erythrocytes
US5364756A (en) * 1990-09-12 1994-11-15 Lifecell Method of cryopreserving a suspension of biological material
US5425951A (en) * 1988-05-18 1995-06-20 Cryopharm Corporation Method of reconstituting lyophilized cells
US5580856A (en) * 1994-07-15 1996-12-03 Prestrelski; Steven J. Formulation of a reconstituted protein, and method and kit for the production thereof
US5690963A (en) * 1995-06-30 1997-11-25 The United States Of America As Represented By The Secretary Of The Navy Freeze dried red blood cells
US5736313A (en) * 1995-10-20 1998-04-07 The United States Of America As Represented By The Secretary Of The Navy Method of lyophilizing platelets by incubation with high carbohydrate concentrations and supercooling prior to freezing
US5750330A (en) * 1996-06-19 1998-05-12 Litron Laboratories Method and composition for lyophilizing red blood cells
US5958670A (en) * 1988-05-18 1999-09-28 Cobe Laboratories, Inc. Method of freezing cells and cell-like materials
US5972592A (en) * 1997-12-29 1999-10-26 Cornell Research Foundation, Inc. Increasing reproductive efficiency of bull semen using fucose or a compound with a fucose moiety
US6007978A (en) * 1988-05-18 1999-12-28 Cobe Laboratories, Inc. Method of freezing cells and cell-like materials
US6040132A (en) * 1996-06-14 2000-03-21 Biostore New Zealand, Ltd. Methods for the lyophilization of living biological materials
US6060233A (en) * 1996-06-14 2000-05-09 Biostore New Zealand, Ltd Methods for the lyophilization of platelets, platelet membranes or erythrocytes
US6274369B1 (en) 1996-02-02 2001-08-14 Invitrogen Corporation Method capable of increasing competency of bacterial cell transformation
US20020009500A1 (en) * 2000-02-10 2002-01-24 Wolkers Willem F. Therapeutic platelets and methods
US6361934B1 (en) * 1997-10-22 2002-03-26 Elizabeth Acton Method and apparatus for cryopreservation
US20020137191A1 (en) * 1996-03-29 2002-09-26 Fredric R. Bloom Method for increasing viability and transformation efficiency of bacteria during storage at low temperatures
US6508983B1 (en) 1999-07-19 2003-01-21 Cobe Cardiovascular Exchanger apparatus and method of manufacture
US20030031998A1 (en) * 1995-06-07 2003-02-13 Prakash Kadkade Cryopreservation of diverse plant cells
US20030035843A1 (en) * 1990-09-12 2003-02-20 Lifecell Corporation, A Delaware Corporation Method for processing and preserving collagen-based tissues for transplantation
US20030091971A1 (en) * 2001-09-14 2003-05-15 Invitrogen Corporation Composition for stabilizing biological materials
US6770478B2 (en) 2000-02-10 2004-08-03 The Regents Of The University Of California Erythrocytic cells and method for preserving cells
US20050031596A1 (en) * 2003-08-06 2005-02-10 Crowe John H. Cells and method for preserving cells
US20050158699A1 (en) * 1995-06-07 2005-07-21 Kadkade Prakash G. Cryopreservation of plant cells
US6960464B2 (en) 1997-02-12 2005-11-01 Invitrogen Corporation Methods for lyophilizing competent cells
US20050277107A1 (en) * 2002-07-26 2005-12-15 Mehmet Toner Systems and methods for cell preservation
US7112576B1 (en) 1999-12-10 2006-09-26 Regents Of The University Of Minnesota Compositions and methods for cryopreservation of peripheral blood lymphocytes
US7358284B2 (en) 1998-06-19 2008-04-15 Lifecell Corporation Particulate acellular tissue matrix
WO2011101796A1 (en) 2010-02-19 2011-08-25 Cadila Pharmaceuticals Limited A pharmaceutical composition of killed cells with substantially retained immunogenicity
US20130230494A1 (en) * 2010-05-25 2013-09-05 The Research Foundation Of State University Of New York Cellular Preparations For Wound Management
US9863699B2 (en) 2014-06-09 2018-01-09 Terumo Bct, Inc. Lyophilization

Families Citing this family (41)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5178884A (en) * 1988-05-18 1993-01-12 Cryopharm Corporation Lyophilized and reconstituted red blood cell compositions
US5059518A (en) * 1988-10-20 1991-10-22 Coulter Corporation Stabilized lyophilized mammalian cells and method of making same
US5153004A (en) * 1989-06-02 1992-10-06 Cryopharm Corporation Freezing and thawing of erythrocytes
JPH05501419A (en) * 1990-05-25 1993-03-18
CA2145418A1 (en) * 1992-09-29 1994-04-14 John S. Patton Pulmonary delivery of active fragments of parathyroid hormone
US20030113273A1 (en) * 1996-06-17 2003-06-19 Patton John S. Methods and compositions for pulmonary delivery of insulin
US6586006B2 (en) 1994-08-04 2003-07-01 Elan Drug Delivery Limited Solid delivery systems for controlled release of molecules incorporated therein and methods of making same
JPH10508744A (en) 1994-09-22 1998-09-02 クオドラント ホールディングス ケンブリッジ リミテッド The composition and manufacturing method thereof for hydration and nutrition during athletic applications
US6290991B1 (en) 1994-12-02 2001-09-18 Quandrant Holdings Cambridge Limited Solid dose delivery vehicle and methods of making same
US6964771B1 (en) 1995-06-07 2005-11-15 Elan Drug Delivery Limited Method for stably incorporating substances within dry, foamed glass matrices
US5762961A (en) * 1996-02-09 1998-06-09 Quadrant Holdings Cambridge Ltd. Rapidly soluble oral solid dosage forms, methods of making same, and compositions thereof
US5958455A (en) * 1996-02-09 1999-09-28 Quadrant Holdings Cambridge Ltd Oral solid dosage forms, methods of making same and compositions thereof
FR2746109B1 (en) * 1996-03-12 1998-04-17 Rhone Poulenc Rorer Sa Medium for conservation biological material
US6632648B1 (en) * 1996-05-14 2003-10-14 Elan Drug Delivery Limited Methods of terminal sterilization of fibrinogen
US6509146B1 (en) 1996-05-29 2003-01-21 Universal Preservation Technologies, Inc. Scalable long-term shelf preservation of sensitive biological solutions and suspensions
US6468782B1 (en) 1996-12-05 2002-10-22 Quadrant Healthcare (Uk) Limited Methods of preserving prokaryotic cells and compositions obtained thereby
JP2001511174A (en) * 1997-02-07 2001-08-07 クアドラント ホールディングス ケンブリッジ リミテッド Dried, methods and compositions for making platelets have a shelf stability
DE19707508C2 (en) * 1997-02-25 2000-05-31 Hemofarm Vrsac A process for the isolation of hemoglobin and the use of the thus isolated hemoglobin
US6071745A (en) 1997-06-27 2000-06-06 Bio-Rad Laboratories Method and formulation for lyophilizing cultured human cells to preserve RNA and DNA contained in cells for use in molecular biology experiments
US20060165606A1 (en) 1997-09-29 2006-07-27 Nektar Therapeutics Pulmonary delivery particles comprising water insoluble or crystalline active agents
US6565885B1 (en) 1997-09-29 2003-05-20 Inhale Therapeutic Systems, Inc. Methods of spray drying pharmaceutical compositions
US5955257A (en) 1997-10-21 1999-09-21 Regents Of The University Of Minnesota Infusible grade short-term cell storage medium for mononuclear cells
CA2332986C (en) 1998-05-26 2009-09-08 Lifecell Corporation Cryopreservation of human red blood cells
US6127177A (en) * 1998-09-11 2000-10-03 Massachusetts Institute Of Technology Controlled reversible poration for preservation of biological materials
DE60016034T2 (en) * 1999-12-21 2005-12-01 Ericson, Daniel G., Rochester Composition for the storage of platelets
ES2525087T5 (en) 2000-05-10 2018-06-28 Novartis Ag phospholipid based powders drug delivery
US8404217B2 (en) 2000-05-10 2013-03-26 Novartis Ag Formulation for pulmonary administration of antifungal agents, and associated methods of manufacture and use
US7871598B1 (en) 2000-05-10 2011-01-18 Novartis Ag Stable metal ion-lipid powdered pharmaceutical compositions for drug delivery and methods of use
US6653062B1 (en) 2000-07-26 2003-11-25 Wisconsin Alumni Research Foundation Preservation and storage medium for biological materials
US7270946B2 (en) * 2002-10-04 2007-09-18 Organ Recovery Systems, Inc. Method for treatment of cellular materials with sugars prior to preservation
US20050095320A1 (en) * 2003-10-29 2005-05-05 Richard Botteri [An Isotonic Sports Drink for Female Athletes Fortified with Iron, Calcium and Essential Vitamins for Use in Rehydration and Nutrition During Execise and Competition]
US20050100636A1 (en) * 2003-11-10 2005-05-12 Megan Botteri [A Low-Calorie Sports Drink For Physically Active Women That is Fortified with Iron, Calcium and Essential Vitamins for Use in Rehydration and Replacing Electrolytes Lost During Periods of Physical Activity]
KR20080103637A (en) * 2007-05-25 2008-11-28 주식회사 알앤엘바이오 Composition for treating of ischemic limb disease comprising stem cells derived from adipose tissue
US8440390B2 (en) * 2009-02-23 2013-05-14 Lifeline Scientific, Inc. Method for ice-free cryopreservation of tissue
US20130008191A1 (en) * 2011-07-07 2013-01-10 Suchak Naresh J Methods for freezing and thawing proteins
US8906668B2 (en) 2012-11-23 2014-12-09 Seres Health, Inc. Synergistic bacterial compositions and methods of production and use thereof
US20160243172A1 (en) 2013-02-04 2016-08-25 Seres Therapeutics, Inc. Compositions and Methods for Inhibition of Pathogenic Bacterial Growth
US10076546B2 (en) 2013-03-15 2018-09-18 Seres Therapeutics, Inc. Network-based microbial compositions and methods
US9956282B2 (en) 2013-12-16 2018-05-01 Seres Therapeutics, Inc. Bacterial compositions and methods of use thereof for treatment of immune system disorders
WO2016086071A8 (en) * 2014-11-24 2016-07-21 Nueon, Inc. Spectrally encoded consumable spectrometer apparatus and methods
US20180192639A1 (en) 2016-12-20 2018-07-12 Tissue Testing Technologies Llc Ice-free preservation of large volume tissue samples for viable, functional tissue banking

Family Cites Families (85)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
FR654692A (en) * 1928-01-04 1929-04-09 Support connection and columns or metal pillars
US2786014A (en) * 1952-09-10 1957-03-19 James L Tullis Platelet preservation
US2875588A (en) * 1953-06-11 1959-03-03 Ohio Commw Eng Co Preservation of red blood cells
US3079919A (en) * 1958-11-10 1963-03-05 Baxter Laboratories Inc Parenteral solution equipment and method
US3228838A (en) * 1959-04-23 1966-01-11 Union Carbide Corp Preservation of biological substances
US3080725A (en) * 1960-08-11 1963-03-12 Union Carbide Corp Method and apparatus for controlled rate cooling and warming of biological substances
US3158283A (en) * 1961-04-24 1964-11-24 Union Carbide Corp Corrugated contained for the low temperature preservation of biological substances
US3344167A (en) * 1962-03-19 1967-09-26 Exxon Research Engineering Co Difluoraminoalkyl nitrates
US3303662A (en) * 1963-03-01 1967-02-14 Union Carbide Corp Process for cell preservation
BE643051A (en) * 1963-05-31 1964-05-15
US3347745A (en) * 1963-12-06 1967-10-17 Union Carbide Corp Process for freezing erythrocytes
FR1413976A (en) * 1964-08-31 1965-10-15 The flexible container of plastics material, especially for serum
US3344617A (en) * 1965-02-25 1967-10-03 Union Carbide Corp Apparatus for the preservation of biological substances
NL6509726A (en) * 1965-07-28 1967-01-30
NL126572C (en) * 1966-03-24
DE1808435B2 (en) * 1967-11-13 1977-06-02 A process for stabilizing erythrocytes
GB1244344A (en) * 1967-11-13 1971-08-25 Abbott Lab Hemagglutination testing procedure employing stabilized and potentiated erythrocytes and a method for their preparation
US3758382A (en) * 1968-07-26 1973-09-11 Us Navy Ve agent process of freezing blood using a hydroxyalkyl starch as cryoprotecti
US3554256A (en) * 1968-11-08 1971-01-12 Dave Champman Goldsmith & Yama Flexible intravenous container
US3677022A (en) * 1969-07-24 1972-07-18 Redeco Method for extending the useful storage period of biological substances such as blood
CA928215A (en) * 1969-08-08 1973-06-12 American Hospital Supply Corporation Process for the cryogenic preservation of blood and erythrocytes and products produced thereby
US3915794A (en) * 1973-02-09 1975-10-28 Rit Rech Ind Therapeut Stabilizing compositions for cell-free viruses and cell-free virus preparations containing them
DE2310964A1 (en) * 1973-03-02 1974-09-05 Schering Ag Antigen or antigen-proteinkonjugatbeschichtete erythrocytenpraeparation
FR2284842B1 (en) * 1974-09-11 1982-07-02 Nestle Sa
DE2532183C3 (en) * 1975-07-18 1982-03-04 Behringwerke Ag, 3550 Marburg, De
CA1055932A (en) * 1975-10-22 1979-06-05 Hematech Inc. Blood substitute based on hemoglobin
US4018911A (en) * 1975-11-10 1977-04-19 The United States Of America As Represented By The Secretary Of The Navy Method for large volume freezing and thawing of packed erythrocytes
DE2551208C3 (en) * 1975-11-14 1981-05-27 Behringwerke Ag, 3550 Marburg, De
US4059967A (en) * 1976-02-19 1977-11-29 The Community Blood Council Of Greater New York, Inc. Process for freezing blood platelets
US4132594A (en) * 1976-06-28 1979-01-02 The United States Of America As Represented By The Administrator Of The National Aeronautics And Space Administration Gas diffusion liquid storage bag and method of use for storing blood
US4131200A (en) * 1976-07-06 1978-12-26 Union Carbide Corporation Thermoplastic blood bag
US4112070A (en) * 1977-06-08 1978-09-05 Research Corporation Blood preservation system
US4320111A (en) * 1977-06-14 1982-03-16 American Home Products Corporation Immunologic compositions methods of preparation and use
US4278198A (en) * 1977-11-17 1981-07-14 Baxter Travenol Laboratories, Inc. Flexible collapsible container with a stiffening member
DE2820603A1 (en) * 1978-05-11 1979-11-15 Studiengesellschaft Kohle Mbh Erythrocytes modified with allo-steric effector(s) - using lipid vesicles, used for increasing oxygen supply to tissues
US4243687A (en) * 1979-01-10 1981-01-06 Leo Kline Freeze-dried natural sour dough starter
ES489149A0 (en) * 1979-03-02 1981-03-16 Unilever Nv A process for the preparation of frozen confections days.
DE2929278C2 (en) * 1979-07-19 1989-03-30 Forschungsgesellschaft Fuer Biomedizinische Technik, 5100 Aachen, De
US4267269A (en) * 1980-02-05 1981-05-12 Baxter Travenol Laboratories, Inc. Red cell storage solution
DE3007913A1 (en) * 1980-03-01 1981-09-17 Fraunhofer Ges Forschung Freezing biological cell material - with freeze-protective agent mixt. contg. extra- and intra-cellular acting components pref. starch derivs.
JPS577419A (en) * 1980-06-17 1982-01-14 Toshiba Kagaku Kogyo Kk Erythrocyte storing solution
US4521975A (en) * 1981-05-04 1985-06-11 Marquest Medical Products, Inc. Lyophilizing and forming biologicals having a predetermined unit dosage
JPS589688A (en) * 1981-07-06 1983-01-20 Toyobo Co Ltd Stable enzymic composition
JPS58131566A (en) * 1982-04-28 1983-08-05 Green Cross Corp:The Lyophilized matter of red blood cell
DE3225408A1 (en) * 1982-07-07 1984-01-12 Biotest Serum Institut Gmbh to suspend aqueous solution and store of cells, in particular erythrocytes
FR2542617B2 (en) * 1982-07-09 1986-06-06 Rgl Transfusion Sanguine Centr protective solution for the conservation of functional cells
US4639513A (en) * 1984-02-02 1987-01-27 Cuno Inc. Intravenously injectable immunoglobulin G (IGG) and method for producing same
US4883459A (en) * 1983-07-29 1989-11-28 Reynaldo Calderon Retrograde perfusion
US4865871A (en) * 1983-08-23 1989-09-12 Board Of Regents The University Of Texas System Method for cryopreparing biological tissue
JPH0411367B2 (en) * 1983-09-10 1992-02-28
US4585735A (en) * 1984-07-19 1986-04-29 American National Red Cross Prolonged storage of red blood cells
US5084377A (en) * 1984-09-19 1992-01-28 Larry Rowan Cryogenic suspension method
FR2581289A1 (en) * 1985-05-06 1986-11-07 Rgl Transfusion Sanguine Centr synthetic solution for prolonged shelf concentrated erythrocyte
GB8604983D0 (en) * 1986-02-28 1986-04-09 Biocompatibles Ltd Protein preservation
JPH0827283B2 (en) * 1986-03-10 1996-03-21 財団法人化学及血清療法研究所 Immunoagglutination composition
US4806343A (en) * 1986-03-13 1989-02-21 University Of Southwestern Louisiana Cryogenic protectant for proteins
US4731330A (en) * 1986-07-01 1988-03-15 Biotrack, Inc. Whole blood control sample
JPS63106562A (en) * 1986-10-23 1988-05-11 Green Cross Corp:The Reagent for reversed passive hemaglutination
US4764463A (en) * 1986-10-30 1988-08-16 The University Of Tennessee Research Corporation Platelet cyropreservation
US4963362A (en) * 1987-08-07 1990-10-16 Regents Of The University Of Minnesota Freeze-dried liposome mixture containing cyclosporin
US4980277A (en) * 1987-10-16 1990-12-25 Cultor Ltd. Cryoprotectant solution and method
US5192553A (en) * 1987-11-12 1993-03-09 Biocyte Corporation Isolation and preservation of fetal and neonatal hematopoietic stem and progenitor cells of the blood and methods of therapeutic use
JPH0769585B2 (en) * 1987-12-14 1995-07-31 富士写真フイルム株式会社 Method for producing photographic silver halide emulsion
US5171661A (en) * 1988-05-18 1992-12-15 Cryopharm Corporation Medium for lyophilization of erythrocytes
US5178884A (en) * 1988-05-18 1993-01-12 Cryopharm Corporation Lyophilized and reconstituted red blood cell compositions
US4900780A (en) * 1988-05-25 1990-02-13 Masonic Medical Research Laboratory Acellular resuscitative fluid
FI80172C (en) * 1988-05-25 1990-04-10 Nokia Mobile Phones Ltd Fm-mellanfrekvenskrets.
DE3817906A1 (en) * 1988-05-26 1989-11-30 Boehringer Mannheim Gmbh The method and container for freeze-dried under sterile conditions
US5045446A (en) * 1988-08-26 1991-09-03 Cryopharm Corporation Lyophilization of cells
US4874690A (en) * 1988-08-26 1989-10-17 Cryopharm Corporation Lyophilization of red blood cells
EP0356258A3 (en) * 1988-08-26 1990-06-06 Cryopharm Corporation Processes for the lyophilization of red blood cells, together with media for lyophilization
US5059518A (en) * 1988-10-20 1991-10-22 Coulter Corporation Stabilized lyophilized mammalian cells and method of making same
US5030560A (en) * 1988-11-04 1991-07-09 Immucor, Inc. Method for drying mammalian cells for use in solid phase immunoassays and articles incorporating same
US4983515A (en) * 1989-02-16 1991-01-08 E. I. Du Pont De Nemours And Company Labeled cryopreserved cells for use as targets in cytotoxicity assays
GB8903593D0 (en) * 1989-02-16 1989-04-05 Pafra Ltd Storage of materials
US5043261A (en) * 1989-04-10 1991-08-27 Cryopharm Corporation Lyophilized and reconstituted red blood cell and hemosome compositions
US5213814A (en) * 1989-04-10 1993-05-25 Cryopharm Corporation Lyophilized and reconstituted blood platelet compositions
US5118792A (en) * 1989-05-10 1992-06-02 Dna Plant Technology Corporation Ice crystal growth suppression polypeptides and method of making
US5153004A (en) * 1989-06-02 1992-10-06 Cryopharm Corporation Freezing and thawing of erythrocytes
GB8920007D0 (en) * 1989-09-05 1989-10-18 Devilbiss Co Spraygun
US4973327A (en) * 1989-11-01 1990-11-27 Cryopharm Blood bag for lyophilization
JPH05501419A (en) * 1990-05-25 1993-03-18
US5145770A (en) * 1990-06-04 1992-09-08 Biosurface Technology, Inc. Cryopreservation of cultured epithelial sheets
JPH05506457A (en) * 1991-02-15 1993-09-22
US5242792A (en) * 1991-02-25 1993-09-07 The United States Of America As Represented By The Secretary Of The Navy Method for the preservation of red blood cells by lyophilization using glycerol or inositol with disaccharides

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
Fujirebio Inc. Chem. Abst. vol. 102, (1985), p. 92530y. *
Fujirebio Inc.-Chem. Abst. vol. 102, (1985), p. 92530y.
Scheiwe et al. Chem. Abst. vol. 98 (1983), p. 14029b. *
Scheiwe et al.-Chem. Abst. vol. 98 (1983), p. 14029b.

Cited By (47)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5958670A (en) * 1988-05-18 1999-09-28 Cobe Laboratories, Inc. Method of freezing cells and cell-like materials
US5340592A (en) * 1988-05-18 1994-08-23 Cobe Laboratories, Inc. Lyophilization of erythrocytes
US6007978A (en) * 1988-05-18 1999-12-28 Cobe Laboratories, Inc. Method of freezing cells and cell-like materials
US5425951A (en) * 1988-05-18 1995-06-20 Cryopharm Corporation Method of reconstituting lyophilized cells
US5648206A (en) * 1988-08-26 1997-07-15 Cobe Laboratories, Inc. Lyophilization of cells
US5336616A (en) * 1990-09-12 1994-08-09 Lifecell Corporation Method for processing and preserving collagen-based tissues for transplantation
US5780295A (en) * 1990-09-12 1998-07-14 Life Cell Corporation Apparatus for cryopreparation, dry stabilization and rehydration of biological suspensions
US8067149B2 (en) 1990-09-12 2011-11-29 Lifecell Corporation Acellular dermal matrix and method of use thereof for grafting
US20060210960A1 (en) * 1990-09-12 2006-09-21 Lifecell Corporation, A Texas Corporation Method for processing and preserving collagen-based tissues for transplantation
US5364756A (en) * 1990-09-12 1994-11-15 Lifecell Method of cryopreserving a suspension of biological material
US20030035843A1 (en) * 1990-09-12 2003-02-20 Lifecell Corporation, A Delaware Corporation Method for processing and preserving collagen-based tissues for transplantation
US6194136B1 (en) 1990-09-12 2001-02-27 Lifecell Corporation Cryoprotective solutions comprising DMSO, PG, 2,3-butanediol,raffinose and PVP
US5800978A (en) * 1992-01-21 1998-09-01 Cobe Laboratories, Inc. Method of freezing cells and cell-like materials
WO1993014191A1 (en) * 1992-01-21 1993-07-22 Cryopharm Corporation Method of freezing cells and cell-like materials
US5580856A (en) * 1994-07-15 1996-12-03 Prestrelski; Steven J. Formulation of a reconstituted protein, and method and kit for the production thereof
US20050158699A1 (en) * 1995-06-07 2005-07-21 Kadkade Prakash G. Cryopreservation of plant cells
US20030031998A1 (en) * 1995-06-07 2003-02-13 Prakash Kadkade Cryopreservation of diverse plant cells
US6753182B1 (en) * 1995-06-07 2004-06-22 Phyton, Inc. Cryopreservation of plant cells
US5690963A (en) * 1995-06-30 1997-11-25 The United States Of America As Represented By The Secretary Of The Navy Freeze dried red blood cells
US5736313A (en) * 1995-10-20 1998-04-07 The United States Of America As Represented By The Secretary Of The Navy Method of lyophilizing platelets by incubation with high carbohydrate concentrations and supercooling prior to freezing
US20040152184A1 (en) * 1996-02-02 2004-08-05 Invitrogen Corporation Method capable of increasing competency of bacterial cell transformation
US6274369B1 (en) 1996-02-02 2001-08-14 Invitrogen Corporation Method capable of increasing competency of bacterial cell transformation
US20100261278A1 (en) * 1996-02-02 2010-10-14 Life Technologies Corporation Method capable of increasing competency of bacterial cell transformation
US6709854B2 (en) 1996-02-02 2004-03-23 Invitrogen Corporation Method capable of increasing competency of bacterial cell transformation
US20020137191A1 (en) * 1996-03-29 2002-09-26 Fredric R. Bloom Method for increasing viability and transformation efficiency of bacteria during storage at low temperatures
US6855494B2 (en) 1996-03-29 2005-02-15 Invitrogen Corporation Method for increasing viability and transformation efficiency of bacteria during storage at low temperatures
US6060233A (en) * 1996-06-14 2000-05-09 Biostore New Zealand, Ltd Methods for the lyophilization of platelets, platelet membranes or erythrocytes
US6040132A (en) * 1996-06-14 2000-03-21 Biostore New Zealand, Ltd. Methods for the lyophilization of living biological materials
US5750330A (en) * 1996-06-19 1998-05-12 Litron Laboratories Method and composition for lyophilizing red blood cells
US6960464B2 (en) 1997-02-12 2005-11-01 Invitrogen Corporation Methods for lyophilizing competent cells
US7648832B2 (en) 1997-02-12 2010-01-19 Life Technologies, Corp. Methods for lyophilizing competent cells
US6361934B1 (en) * 1997-10-22 2002-03-26 Elizabeth Acton Method and apparatus for cryopreservation
US5972592A (en) * 1997-12-29 1999-10-26 Cornell Research Foundation, Inc. Increasing reproductive efficiency of bull semen using fucose or a compound with a fucose moiety
US7358284B2 (en) 1998-06-19 2008-04-15 Lifecell Corporation Particulate acellular tissue matrix
US6508983B1 (en) 1999-07-19 2003-01-21 Cobe Cardiovascular Exchanger apparatus and method of manufacture
US7112576B1 (en) 1999-12-10 2006-09-26 Regents Of The University Of Minnesota Compositions and methods for cryopreservation of peripheral blood lymphocytes
US20040175374A1 (en) * 2000-02-10 2004-09-09 Wolkers Williem F. Therapeutic platelets and methods
US20020009500A1 (en) * 2000-02-10 2002-01-24 Wolkers Willem F. Therapeutic platelets and methods
US6723497B2 (en) 2000-02-10 2004-04-20 The Regents Of The University Of California Therapeutic platelets and methods
US6770478B2 (en) 2000-02-10 2004-08-03 The Regents Of The University Of California Erythrocytic cells and method for preserving cells
US20030091971A1 (en) * 2001-09-14 2003-05-15 Invitrogen Corporation Composition for stabilizing biological materials
US20050277107A1 (en) * 2002-07-26 2005-12-15 Mehmet Toner Systems and methods for cell preservation
US20050031596A1 (en) * 2003-08-06 2005-02-10 Crowe John H. Cells and method for preserving cells
WO2011101796A1 (en) 2010-02-19 2011-08-25 Cadila Pharmaceuticals Limited A pharmaceutical composition of killed cells with substantially retained immunogenicity
CN102753975A (en) * 2010-02-19 2012-10-24 卡帝拉药物有限公司 A pharmaceutical composition of killed cells with substantially retained immunogenicity
US20130230494A1 (en) * 2010-05-25 2013-09-05 The Research Foundation Of State University Of New York Cellular Preparations For Wound Management
US9863699B2 (en) 2014-06-09 2018-01-09 Terumo Bct, Inc. Lyophilization

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